[Conditionally pathogenic microorganisms in patients with bisphosphonate jaw osteonecrosis].

The objective of the study was to define treatment strategy in cases of facial bones bisphosphonate induced osteonecrosis based on the study of the role of conditionally pathogenic oral microorganisms. Three typical clinical cases of bisphosphonate osteonecrosis of the facial bones were analyzed and 15 conditionally pathogenic oral microorganisms were identified in these patients using real-time PCR in saliva, wound and bone samples. A comparative analysis was carried out with purulent-inflammatory diseases of maxillofacial area. The study results proved an important role of conditionally pathogenic microorganisms of the oral cavity in the development of osteonecrosis of the facial bones. Wide range of bacterial species was identified in osteonecrosis of the facial bones patients. While bone tissue is most exposed to microbial communities, surgical treatment results in effective rehabilitation for a long period.

[Detection of Staphylococcus aureus toxins using immuno-PCR].

A highly sensitive test system, based on the method of immuno-PCR, was developed for the detection of two staphylococcal toxins: enterotoxin A (SEA) and toxic shock syndrome toxin (TSST). A key element of the developed systems was obtaining supramolecular complexes of bisbiotinylated oligodeoxynucleotides and streptavidin, which were used as DNA tags. Specificity studies showed no cross-reactivity when determining SEA and TSST. The sensitivity of detection of these toxins in the culture supernatants S. aureus was not less than 10 pg/mL.

[Immunogenetic aspects of early rheumatoid arthritis].

The study is aimed to investigate the distribution of alleles of HLA-DRB1 gene in patients with early rheumatoid arthritis and healthy individuals in Russian population, and evaluate their significance as molecular genetic markers of rheumatoid arthritis predisposition and protection. The association between alleles of HLA-DRB1 genes, antibodies to cyclic citrullinated peptides and IgM rheumatoid factor was also studied. Low and high resolution HLA-DRB1 genotyping were compared. In the cohort of patients with early rheumatoid arthritis, the alleles of HLA-DRB1 gene were found to be markers of rheumatoid arthritis protection/risk, especially in the homozygous state. They determined production of antibodies to cyclic citrullinated peptides but were not associated with rheumatoid factor IgM levels. These findings support different autoimmune mechanisms of rheumatoid arthritis pathogenesis.

[Immunogenetics of type 1 diabetes mellitus--from fundamental ideas to medical practice].

The review of studies of Russian researchers on theoretical and practical aspects of genetic predisposition to type 1 diabetes associated with immunity: HLA and not HLA genes. Most important for practical public health outcomes are evidence that HLA-genetic predisposition to type 1 diabetes is associated with the DRB1-genotype, consisting entirely of variants DRB1-genes associated with the development of T1D. It was also established that CTLA4 gene has an independent predictive value for T1D.

[The development and testing of a molecular biological test system for DNA detection of anthrax pathogen by real-time polymerase chain reaction assay].

The paper presents the results of the development and testing of a molecular biological test system for DNA detection of anthrax pathogen (Bacillus anthracis) by real-time polymerase chain reaction assay. The test system has shown high sensitivity, specificity, and reproducibility of results of analysis, as exemplified by aqueous suspensions of daily agar cultures of Bacillus anthracis strains, related and heterologous species of microorganisms, and clinical materials of experimental animals. There is evidence for the persistence of the basic characteristics of the test system when stored at 22 +/- 2 degrees C for 12 months.

[PCR technique for prenatal diagnosis of fetal rhesus factor from peripheral maternal blood].

A procedure is proposed for the noninvasive determination of fetal rhesus factor in rhesus-negative pregnant women, which is based on the detection of the RHD gene in peripheral maternal blood by a PCR technique. The studies have shown the high sensitivity and specificity of determination of rhesus factor in a fetus at more than 15 weeks gestation. The technique does not require the use of such invasive and pregnancy-threatening procedures, as amniocentesis, cordocentesis or chorion biopsy. Furthermore, determination of fetal blood rhesus factor in rhesus-negative patients makes it possible to reduce expenses on the management of pregnancy, to avoid multiple determination of antibody rhesus and prevention of rhesus immunization, and to timely initiate therapeutic and preventive measures.

[A new method for quantitative estimation of the number of virus particles].

[In the present work virus particles of live mumps virus vaccine widely used for vaccination in Russia have been detected and visualized by atomic force microscopy. For quantitative estimation of the number of observed virus particles the special method has been proposed. The presence of protein component of the virus in vaccine was tested by ELISA and dot-blot analysis. Using quantitative real-time PCR assay the number of copies of viral RNA was estimated. The results of quantitative estimation obtained by real-time PCR corresponded with atomic force microscopy data.

[The use of achievements in human molecular immunogenetics in the management of type 1 diabetes mellitus].

New original data are presented on the use of achievements in human molecular immunogenetics in the management of type 1 diabetes mellitus. They include materials allowing for the prediction of the development of the disease at the population, family, and individual levels along with novel approaches to its radical treatment by the reconstitution of the lost glucose tolerance. The reported data may find wide application in current clinical practice. They open up new prospects for the enhancement of efficacy of prognosis, diagnosis, and treatment of type 1 diabetes mellitus and other autoimmune diseases.

[The use of polymerase chain reaction for detection of the agents of glanders and melioidosis using experimental infection].

Glanders and melioidosis are severe infectious diseases of people and animals. The causative agents of these infections refer to the potential agents of bioterrorism of group B. In this work the possibility of use of flagellin-based primers for the identification of B. mallei and B. pseudomallei and for diagnosis of experimental glanders and melioidosis was studied. The obtained results permit to make a conclusion that PCR using the developed primers may be recommended for the incorporation in the scheme of laboratory diagnosis of glanders and melioidosis both for the identification of clean cultures and in experimental clinical material.

[Typing of group A streptococci isolated from urban population of different social status and age].

Pulse-electrophoresis, sequencing of emm genes coding protein M and PCR analysis of speA, speB, and speC genes were used for characterization of group A streptococci (GAS) isolated in different years in Moscow and Tuapse mostly from children and military staff. It has been shown that epidemic process of streptococcal infection caused by GAS in Moscow is based on circulation of many independent clones of Streptococcus pyogenes. Obtained data on complex typing of S. pyogenes would be useful for study of molecular epidemiology of diseases caused by GAS and improvement of epidemiologic surveillance.

[Accuracy of a real-time polymerase-chain-reaction assay for a quantitative estimation of genetically modified sources in food products].

The accuracy of a real-time polymerase-chain-reaction assay for genetically modified sources in food products was determined using two official test systems (kits) of primers and samples. These kits were recommended by the Federal Center of State Sanitary and Epidemiological Surveillance (Russian Ministry of Health) and the European Commission. We used the following three models of thermocyclers: iCycler iQ (BioRad, United States), Rotor-Gene 3000 (Corbett Research, Australia), and DT-322 (DNA-Technology, Russia). Studies of samples that contained 1% genetically modified sources showed that the error of a quantitative assay for genetically modified sources in food products corresponds to 20-30% and does not depend on the kit type and the thermocycler model used.

[Real-time PCR: approaches to data analysis (a review)].

The registration of the accumulation of polymerase chain reaction (PCR) products in the course of amplification (real-time PCR) requires specific equipment, i.e., detecting amplifiers capable of recording the level of fluorescence in the reaction tube during amplicon formation. By the time the reaction is completed, researchers obtain DNA accumulation graphs. The review discusses the most promising algorithms of analysis of real-time PCR curves and possible errors, whether caused by the software used or the operators' mistakes. The data included will assist researchers in understanding the features of the method to obtain more reliable results.

[Use of PCR for identification of Burkholderia mallei]. / Ispol'zovanie PTsR dlia identifikatsii Burkholderia mallei.

Stimuli of glanders belong to the potential agents of biological terror. The possibility to use various primers in the identification of B. mallei was investigated and the significance of polymerase chain reaction (PCR) was defined within the scheme of laboratory glanders diagnosis in the offered paper. The constructed amplifying test-systems can be used to detect the glanders both in the environmental objects contaminated with B. mallei and in experimental clinical material.

[Frequency of genes speA, speB, and speC in Streptococcus pyogenes strains and the identification of the infective agent by polymerase chain reaction]. / Chastota vstrechaemosti genov speABC v shtammakh Streptococcus pyogenes i identifikatsia vosbuditelia s pomoshch'iu PTSR.

In cultures of S. pyogenes isolated from patients and carriers in different territories of the Russian Federation the genes of erythorogenic toxins A, B and C (speA, speB and specC) were detected. The possibility of the identification of S. pyogenes by means of PCR on the basis of primers to erythrogenic toxin B was determined. Gene speB was detected in all S. pyogenes cultures under study and proved to be species specific. Genes speA and speC were detected, respectively, in 29.4% and 9.35% of the S. pyogenes cultures under study. A test system for the identification of S. pyogenes on the basis of primers to gene speB was developed. The prospects for the detection of genes speA and speC for intraspecific typing of this infective agent were evaluated.

[The risk factors for the development of multiple sclerosis in the Moscow population. II. The combination of exogenous and hereditary factors]. / Faktory riska razvitiia rasseiannogo skleroza v moskovskoi populiatsii. II. Sochetaniia ékzogennykh i nasledstvennykh faktorov.

Simultaneous study of both external and hereditary risk factors of the development of multiple sclerosis (MS) was carried out in Moscow population. As a result of the genotyping of HLA-DR locus on the chromosome 6, there were found markers of the predisposition to MS: by HLA-DR: DR2(15) way, especially, in homozygous heredity and in the absence of such specificity--alleles of DR3 group. The data were confirmed concerning both the elevation of frequency of the alleles of tumor necrosis factor (TNF)a1 and alpha 2 and the decrease of frequency of TNF-alpha 7 allele in MS. There was also confirmed association of the disease with the external factors, such as "predominance of the meat nutrition at the age under 15 years" and "tonsillitis at the age under 15 years". There were two combinations of the external and hereditary factors, when the power of the association increased significantly: "predominance of meat nutrition at the age under 15 years" and DR2(15) and "tonsillitis at the age under 15 years" and DR3. When genetic factors were excluded, the relation of MS with the external factors was considerably reduced; when the external factors were excluded, the association of MS with genetic markers became statistically insignificant. A combination of these external factors, which were the sources for the additional antigenic immunostimulation, with certain genetic factors in DR region suggested the presence of the genetically determined peculiarities of the composition of antigen presentation HLA-DR in the individuals with DR2(15) and DR3. There was no statistically confirmed presence of a combination of the external factors with TNF-alpha alleles which increased additionally MS risk. The presence of immunostimulating antigen-containing external factors may be significant for the influence of TNF-alpha in less degree than for the realization of predisposition in form of the certain composition of HLA molecules.

[Genetic predisposition factors for multiple sclerosis (based on the data from genotyping patients of the Russian ethnic group)]. / Faktory geneticheskoi predraspolozhennosti k rasseiannomu sklerozu (po dannym genotipirovaniia bol'nykh russkoi étnicheskoi gruppy).

Patients' genotyping of Russian ethnic group with multiple sclerosis (MS) was performed for the first time in two loci of the main complex of histocompatibility: in DR HLA class II (gene DRB1) and in the locus of tumor necrosis factors (TNF). There was no difference in the incidence of alleles groups which correlated with some specificity of DR. Meanwhile TNF-a1 and TNF-a9 alleles were encountered significantly more frequently in patients and TNF-a7 in control group. When all the patients and controls examined were divided into groups in dependence on combination of DR and TNF it was found that relations observed between MS and TNF-a7 and TNF-a9 alleles were displayed much more in individuals which carried alleles of gene DRB1, corresponding to DR15 specificity. These are alleles which are known as the main risk factor of MS in Caucasians. The patients with "protective" TNF-a7 allele were characterized by more favorable course of the disease. Thus highly significant genetic markers were revealed for the first time in region of TNF genes which were associated with increased or decreased risk of MS development at least in Russian ethnic group. There was also possibility of their interaction with group of DR15 alleles of DRB1 gene. One of the markers revealed (TNF-a7) occurred to be bound both with decreased risk of MS and with favorable clinical course, which was observed for genetic markers of MS for the first time. Manifestation of one or another property of TNF-a7 marker depends on the presence of alleles of DRB1 gene which corresponds to DR15 specificity.